Compensatory Feto-Placental Upregulation of the Nitric Oxide System during Fetal Growth Restriction

Background: Fetal Growth Restriction is often associated with a feto-placental vascular dysfunction conceivably involving endothelial cells. Our study aimed to verify this pathogenic role for feto-placental endothelial cells and, coincidentally, demonstrate any abnormality in the nitric oxide system. Methods: Prenatal assessment of feto-placental vascular function was combined with measurement of nitric oxide (in the form of S-nitrosohemoglobin) and its nitrite byproduct, and of the endogenous nitric oxide synthase inhibitor asymmetric dimethylarginine. Umbilical vein endothelial cells were also harvested to determine their gene profile. The study comprised term pregnancies with normal (n = 40) or small-for-gestational-age (n = 20) newborns, small-for-gestational-age preterm pregnancies (n = 15), and bi-chorial, bi-amniotic twin pregnancies with discordant fetal growth (n = 12). Results: Umbilical blood nitrite (p<0.001) and S-nitrosohemoglobin (p = 0.02) rose with fetal growth restriction while asymmetric dimethylarginine decreased (p = 0.003). Nitrite rise coincided with an abnormal Doppler profile from umbilical arteries. Fetal growth restriction umbilical vein endothelial cells produced more nitrite and also exhibited reciprocal changes in vasodilator (upwards) and vasoconstrictor (downwards) transcripts. Elevation in blood nitrite and S-nitrosohemoglobin persisted postnatally in the fetal growth restriction offspring. Conclusion: Fetal growth restriction is typified by increased nitric oxide production during pregnancy and after birth. This response is viewed as an adaptative event to sustain placental blood flow. However, its occurrence may modify the endothelial phenotype and may ultimately represent an element of risk for cardiovascular disease in adult life.Fil: Pisaneschi, Silvia. Università degli Studi di Pisa; Italia. Scuola Superiore Sant’Anna; ItaliaFil: Strigini, Francesca A. L.. Università degli Studi di Pisa; ItaliaFil: Sanchez, Angel Matias. Università degli Studi di Pisa; Italia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Begliuomini, Silvia. Università degli Studi di Pisa; ItaliaFil: Casarosa, Elena. Università degli Studi di Pisa; ItaliaFil: Ripoli, Andrea. National Research Council. Institute of Clinical Physiology, ; ItaliaFil: Ghirri, Paolo. Università degli Studi di Pisa; ItaliaFil: Boldrini, Antonio. Università degli Studi di Pisa; ItaliaFil: Fink, Bruno. Noxygen Science Transfer and Diagnostics; AlemaniaFil: Genazzani, Andrea R.. Università degli Studi di Pisa; ItaliaFil: Coceani, Flavio. Scuola Superiore Sant’Anna; ItaliaFil: Simoncini, Tommaso. Università degli Studi di Pisa; Itali

Selective effect of chlormadinone acetate on brain allopregnanolone and opioids content

Background: Synthetic progestins may have different biological actions depending on the target tissue, the dose administered or the coadministration of an estrogen molecule. The purpose of the present study was to evaluate the neuroendocrine effect of chlormadinone acetate (CMA) administration, analyzing the brain content of allopregnanolone (ALLO), an endogenous neurosteroid γ-aminobutyric acid agonist with anxiolytic properties, and the brain level of β-endorphin (β-END), an endogenous opioid implicated in pain mechanism, emotional state and autonomic control. Study Design: Seven groups of Wistar ovariectomized (OVX) rats received one of the following treatments: oral CMA at a dose of 0.1, 0.5 or 1 mg/kg per day; estradiol valerate (E2V) at a dose of 0.05 mg/kg per day; CMA plus E2V (CMA 0.1 or 0.5 or 1 mg/kg per day + E2V 0.05 mg/kg per day) for 14 days. One group of fertile rats and one group of OVX rats were used as controls. Results: CMA increased ALLO content in the hippocampus and, when it was administered with E2V, also in the hypothalamus and anterior pituitary, evidence of a synergic effect with estrogens only in selective brain areas. β-END content increased in the neurointermediate lobe and anterior pituitary after CMA administration, and it did not antagonize the positive, estrogen-induced increase of β-END level. Conclusion: CMA is effective in increasing ALLO and β-END in selective brain areas showing a specific pattern of interaction with brain function, different compared to progesterone or to other synthetic progestins. In particular, CMA action on part of the limbic system (hippocampus and hypothalamus) and on the anterior pituitary support the hypothesis that this progestin might affect cognitive function, emotional state and autonomic control. © 2009 Elsevier Inc. All rights reserved

Sexually dimorphic effects of testosterone administration on brain allopregnanolone in gonadectomized rats

Clinical and biological evidences have shown a wide range of neuroactive effects of testosterone administration

Time-related change in peripheral blood levels of NO₂ and NOHb in AGA and SGA newborns.

<p>Blood levels of NO₂ (<b>A, C</b>) and NOHb (<b>B, D</b>) in relation to BW centile (<b>A, B</b>) and prenatal Doppler finding (<b>C, D</b>) at different time intervals after birth. BW centile: 75^th–25^th (black circles); <25^th–10^th (black and white circles); <10^th–3^rd (white circles); <3^rd (white squares). Doppler finding for term (T) and preterm (PT) pregnancy: normal (white circles) and abnormal (black circles) with each point representing a single patient. *p<0.05; **p<0.01; ***p<0.001 for 24- and 72- h intervals relative to time zero. Note that, in the aggregate, NO₂ and NOHb values for 75^th–25^th centile are significantly different from the other groups (p<0.001) at the 24- and 72-h time intervals, the only exception being the NOHb value for the <3^rd centile at the 72-h mark (p = 0.068).</p

Prenatal umbilical artery Doppler velocimetry vs. NO function in fetuses with normal or restricted growth.

<p>Blood levels of NO₂ (<b>A</b>), NOHb (<b>B</b>), and ADMA (<b>C)</b> for umbilical vein and artery with normal (N) vs. abnormal (Abn) Doppler velocimetry in term (T) or preterm (PT) pregnancies. Note that number of patients is low for certain groups (n = 1 or 2; see columns without SD bar), while n = 3–18 for the remainder. HUVEC from pregnancies with normal vs. abnormal Doppler: eNOS activity with attendant NO₂ formation (<b>D</b>) (n = 3–4) and eNOS protein expression (<b>E</b>) (n = 2). *p<0.05; **p<0.01 vs. normal Doppler.</p

NO₂, NOHb and ADMA levels in umbilical blood from pregnancies with normal or restricted fetal growth.

<p>Umbilical vein and artery levels of NO₂ (<b>A, B</b>), NOHb (<b>C, D</b>) and ADMA (<b>E, F</b>) at different BW centiles: 75^th–25^th (black circles); <25^th–10^th (black and white circles); <10^th–3^rd (white circles); <3^rd (white squares). Dashed line presents the exponential or linear relationship between these variable and BW. *p<0.05; **p<0.01; ***p<0.001.</p

NO₂ and NOHb levels in umbilical blood from twins with discordant intrauterine growth.

<p>NO₂ (<b>A, B</b>) and NOHb (<b>C, D</b>) in umbilical vein and artery blood. NT: twin with normal growth; ST: twin with restricted growth. Note that points for each set of twins are connected by a line. *p<0.05; **p<0.01 vs. normal twin.</p

Gene profile in HUVEC from SGA vs. AGA pregnancies.

<p>Comparison of gene expression at 75^th-25^th (black squares) vs. <3^rd (white squares) BW centile. Figure reports transcripts being affected by fetal growth restriction that relate to hemodynamic control (<b>A</b>), angiogenesis (<b>B</b>), extracellular matrix turnover (<b>C</b>) and inflammatory/adhesion process (<b>D</b>)<b>.</b> Values are expressed as the intensity ratio between test and reference genes (for details, see Methods). Note that the total gene cohort under examination is given in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0045294#pone.0045294.s005" target="_blank">Table S4</a>. *p<0.05; **p<0.01; ***p<0.001 for <3^rd vs. 75^th–25^th BW centile.</p